Nucleoside Q was found in the first position of anticodon in some transfer ribonucleic acids (tRNAs). Total synthesis of the nucleoside Q and its diastereomer made it possible to determine relative and absolute configurations of nucleoside Q. An unusual posttranscriptional modification mechanism in the biosynthesis of nucleoside Q was found by Nishimura et al. by using our synthetic nucleoside Q derivatives. A deficiency of Q in tRNAs is related to tumor growth. Some cancer cells contain tRNAs having G in place of Q ; Q base administered can be incorporated into these tRNAs. Two anticancer antibiotics having such 7-deazapurine nucleus as cadeguomycin and kanagawamicin were found and the former was synthesized.
New saponins, named gypenosides XXVII (1), XXVIII (2), XXXVIII (3), XXXIX (4), XL (5), XLI (6) and LV (7) were isolated from the aerial parts of Gynostemma pentaphyllum MAKINO. collected in Hyogo pref. Structures of 1-6 and 7 were established as 3-O-β-sophoroside of dammer-24-ene-3β, 19, 20 (S)-triol, dammar-24-ene-19-oxo-3β, 20 (S)-diol, dammar-24-ene-3β, 12β, 19, 20 (S)-tetraol, dammar-24-ene-3β, 12β, 19, 20 (R)-tetraol, dammar-24-ene-19-oxo-3β, 12β, 20 (R)-triol, dammar-24-ene-3β, 19, 20 (R)-triol and as 3-O-β-D-glucopyranosyl-20 (S)-[β-xylopyranosyl (1→6)-β-D-glucopyranoside of dammar-24-ene-3β, 19, 20 (S)-triol, respectively by means of carbon-13 nuclear magnetic resonance spectroscopy.
Since proliferation of Propionibacterium is thought to play a significant role in inflammation of acne vulgaris, the antibacterial activities of several crude drugs to this bacteria were examined by agar diffusion method using paper disc, and it was found that the ethanol extract of flower buds of Sophora japonica L. shows a remarkable antibacterial activity against Propionibacterium acnes, P. avidum, Staphylococcus aureus under weak acidic conditions. After purification of active component (s) of the extract, the activity was found to be attributed to the result of the interaction caused by such three components as quercetin (I), rutin (II) and isorhamnetin-3-rutinoside (III). When I, II and III were applied singly, only I shows very weak activity, but others do not show at all. By mixing I and II, or I and III, 50% or 70% activity of the ethanol extract appeared, respectively. Moreover, after mixing the three flavonoids, the full activity of the extract was recovered. A small amount of kaempferol-3-rutinoside was also obtained from this extract. This compound was found to have a similar effect to that III has, though weaker than the latter. The examination of this interaction by use of liquid culture system, showed that the antibacterial activity of the system increased with an increase in the solubility of I caused by II. That is to say, the true principle of the activity is I, and by the addition of II to it, the solubility of I increased to increase apparent activity. A similar increase in activity by the addition of III is thought to be based on the same mechanism as II.
A sensitive, rapid and reproducible high performance liquid chromatographic (HPLC) determination of ephedrine and glycyrrhizin in human breast milk is described. Chromatography was performed on a C-18 bonded stationary phase with 2% acetic acidacetomtrile (2 : 1) as mobile phase, phenytoin as internal standard and ultraviolet detector. Solvent extraction of the drugs are not necessary ; however, one has only to precipitate the breast milk proteins. The peak-height ratio of ephedrine and glycyrrhizin to phenytoin is linearly related to amounts up to 30μg of ephedrine and 10μg of glycyrrhizin per injection. The proposed method was applied to the human breast milk of the patients receiving "Kakkonto extract granules"(Tsumura Jyuntendo Co., Ltd.).
Fluorescent intensity of lumogallion, a fluorescent complex by the addition of aluminium ion after association with primary, secondary and tertiary n-alkylamines was compared with that of calcon. In the result, lumogallion formed a well extractable complex with tertiary amines but merely produced a complex with secondary amines and not produced a complex with primary amines. This method was applied to the determination of local anesthetics and psychotropic agents. Carpipramine, opipramol, benactyzine, chlorpromazine, procaine and lidocaine could be determined in the range of 2×10-8-10-9 mol/ml sample (ca. 8×10-10-10-11 mol/ml measurement solution). Primary and secondary amines at least less than equal molar in the sample did not interfer the determination. Recoveries of carpipramine and chlorpromazine in milk were 99 to 103%.
Influence and mechanisms of four immunostimulants, Maycobacterium bovis (BCG), Streptococcus preparation (OK-432), Corynebacterium parvum (C. parvum), Lactobacillus casei YIT 9018 (LC 9018) and Lactobacillus fermentum YIT 0159 (LF), which had no immuno stimulating activity, on the activity of hepatic drug-metabolizing enzyme and on the activity of glucose-6-phosphate dehydrogenase (G6PD) and on the oxygen radical production by fixed macrophages in the liver of BALB/c mouse were studied. The levels of cytochrome P-450 and the activities of aminopyrine demethylase (APD), nicotinamide adenine dinucleotide phosphate-cytocrome c (NADPH-cytochrome c) reductase significantly decreased after the i. v. injection of BCG, LC 9018, OK-432, C. parvum and LF compared to the saline-injected control mice, but the level of cytochrome b5 was not altered with these treatments. On the other hand, G6PD activity in the liver more remarkably increased with these immunostimulants than the saline i. v. injected mice, and oxygen radical production by fixed macrophages in the liver significantly increased after i. v. injection of LC 9018 and C. parvum. It was suggested that the ability to reduce the enzyme activities of APD and NADPH-cytochrome c reductase and the levels of cytochrome P-450 and cytochrome b5 of the liver has no correlation with tumoricidal and bactericidal activity of these bacterial immunostimulants.
During our studies on the active moiety of anti-ulcerogenic protein in rabbit skin tissues infected with vaccinia virus, we obtained a tetrapeptide, Gly-Ser-His-Lys (GSHL), as the smallest essential residue. The action of this peptide widely covered against various experimental ulcers in rats induced by pylorus-ligation, restraint plus water immersionstress (minimum effective dose, mg/kg, i. v. ; 5.0), aspirin (5.0), histamine (5.0), cortisone (2.5/d), cysteamine (10) and acetic acid (1.0/d). In the respect of minimum effective dose, the effect of GSHL was 5-20 fold stronger than that of cimetidine, except for acetic acidinduced ulcers. Histological finding of acetic acid-induced ulcers revealed that GSHL treatment shortened an apparent defect of mucosa, a true defect of mucosa and rupture of muscaliris mucosae, and that it increased healing index and mucosal regeneration index. The peptide significantly decreased serum gastrin level, increased mucosal and stromal hexosamine contents and also increased mucosal blood flow in stress-treated rats. It significantly inhibited gastric secretion in pylorus-ligated rats. The mechanism of the action of GSHL appears to be due to the restorative effect against defected mucosa and anti-secretory effect.
The relationship between the toxicity and anti-cholinesterase action of 4-methylthiophenyl dipropyl phosphate (propaphos) was investigated. Oral administration of propaphos induced tremor or convulsion at a dose above 50 mg/kg in mice and at higher dose the lethal effect of propaphos was observed dose-dependently. The LD50 was 104 (96-113) mg/kg. Propaphos inhibited blood acetylcholinesterase activity of mice at low dose (25 mg/kg), while brain acetylcholinesterase activity was not affected by the dose but reduced by large dose of propaphos (100-150 mg/kg) markedly. These results suggested that the lethal action of propaphos may be closely associated with the inhibition of brain acetylcholinesterase activity. Further, propaphos was tested on general pharmacological actions in mice, rats, guinea pigs and rabbits, and the results were as follows : (1) Propaphos did not show remarkable central actions in mice when administered orally. (2) In in vitro experiments, propaphos showed non-specific inhibitory effect on the contraction induced by several agents in rat diaphragm and atria, ileum and trachea smooth muscle of guinea pig.
The effects of water extracts of ten Artemisia plants [1, A. japonica THUNB. (spike, leaf) ; 2, A. apiacea HANCE (spike, leaf) ; 3, A. annua L. (spike, leaf) ; 4, A. princeps PAMP. (spike, leaf) ; 5, A. capillaris THUNB. (spike, leaf) ; 6, A. iwayomogi KITAM. (spike, leaf) ; 7, A. keiskeana MIQ. (spike) ; 8, A. feddei LAV. et VAN. (spike) ; 9, A. stolonifera (MAXIM.) KOMAR. (spike) ; 10, A. stelleriana BESS. (spike)] on bile flow, and biliary secretion of bile acids, cholesterol and phospholipids were examined in Wistar rats. Most of these extracts increased bile flow in 30 min after administration, and 1 (leaf) did after 30 min. The increased bile flow in 2 (spike), 3 (spike), 5 (spike), 6 (spike, leaf) and 7 (spike) gradually decreased thereafter, but that in 1 (leaf), 3 (leaf), 4 (spike, leaf), 5 (leaf), 8 (spike) and 9 (spike) remained at higher levels even in 120 or 150 min. Biliary secretion of bile acids, cholesterol and phospholipids decreased after administration of most of these extracts. These changes were not related to those in the bile flow.
The constituents of the fruits of Ligustrum obtusifolium SIEB. et ZUCC. were examined. Ligustroside, 10-hydroxyligustroside, (+)-pinoresinol-β-D-glucopyranoside, (-)-olivil-4"-β-D-glucopyranoside and acteoside were isolated from the methanolic extract. In addition, oleoside 7-methyl ester, 10-hydroxyoleoside 7-methyl ester and methyl α-D-galactopyranoside were obtained, which were found to be artifacts formed during the extraction with MeOH.
The antioxidative effects of about one hundred methanol extracts of crude drugs were examined on the air oxidation of linoleic acid. Several methanol extracts had a moderately strong antioxidative effect. Especially, the materials of spices and aromatic crude drugs such as Zingiberis Rhizoma, Magnolicae Cortex, and Curcumae Rhizoma possessed a relatively strong activity.
Cationic surfactants such as cetyltrimethylammonium chloride and benzalkonium chloride were proved to be quantitatively analyzed by extracting as their ion-paired complexes of lumogallion, which is used as a reagent for the fluorescent determination of some metal ions. Thus, these complexes were most conveniently extracted with dichloromethane, and quantitatively analyzed by fluorometry upon addition of Al8+. Lumogallion was invariably extracted with these quarternary ammonium surfactants at pH range from 7.5 to 3.0, whereas it was negligibly extracted above neutral pH region in the presence of some diverse materials such as primary, secondary, and tertiary amines. Therefore, 7.3 was chosen as an optical pH in the extraction procedure for the quantitative analysis. The present analysis was not diversed in the presence of some metals. The calibration curves showed a good linearity over the range of 0.25-2.5μg/sample (ca. 0.01-0.1μg/ml). The coefficients of variation for the present method were 3.3 to 5.7%.
The automatic operation of the programmed flow preparation and analysis of the component in biological fluids was examined. Three steps including high efficiency direct column extraction using fine diatomaceous earth granules and diethyl ether, on-line evaporation of the solvent in the micro amount of an extract and a switched columninjection into the liquid chromatograph were linked together by multi-channel valves and PTFE tubes. According to the program, the time-lag required for the unit operation is determined by preliminary experiments and the pumps and valves are controlled by an electric sequencer. This system was applied to the analysis of human urine caffeine content.
Effects of serum cholinesterase activity on muscle relaxation caused by succinylcholine were examined in rats. By using acetylthiocholine as substrate, serum cholinesterase activity was estimated colorimetrically. For pharmacologic response, the twitch hight of tibialis anterior muscle caused by electrical stimulation of sciatic nerve was determined. Correlations between serum cholinesterase activity and maximal relaxation or between serum cholinesterase activity and recovery time to 10% response were not significant. A negative correlation, though not significant, was observed between serum cholinesterase activity and area under response time curve with higher dose level (0.3 and 0.4 mg/kg). It was concluded that interindividual variation of serum cholinesterase activity has no significant effect on pharmacologic response to succinylcholine in rats.
Heating and irradiation with room-fluorescent light as well as sunlight had little effect on the stability of a sterilized aqueous solution of sodium nitrite. Nitrite ion in an acidic medium became expectedly unstable with a considerable reduction of the half-life. After the intravenous injection of sodium nitrite solution to rabbits, the blood concentrations of nitrite ion decreased rapidly according to a first-order manner. The hydralazine-high performance liquid chromatography method employed here was proved to be useful for the determination of nitrite ion in biological and environmental samples.
To cause clioquinol intoxication to rabbits, a fine clioquinol suspension for injection prepared with polysorbate 80 was administered intravenously and plasma concentration of clioquinol and its metabolites, clioquinol glucuronide and sulfate, and tissue concentration of clioquinol were determined. Mean plasma concentration of clioquinol 2 min after single administration of 20 mg/kg dose of clioquinol was 165 nmol/ml and only 1.4 nmol/ml 3 h later. Plasma concentrations of the two metabolites reached maximum values within 5-15 min after administration and thereafter, decreased rapidly. Clioquinol concentration was higher in the kidney and liver than those in the other tissues. In repeated administration, plasma concentrations of clioquinol and the metabolites after the initial administration were approximately similar to those in the subsequent administration. In repeated administration of 20 mg/kg/d dose of clioquinol, 2 of 10 rabbits developed ataxia on the hind legs on the 28th and 35th days, respectively, after starting administration. Pathologically, the peripheral nerves showed an overt degeneration. In the nervous system, clioquinol concentration was higher in the sciatic nerve than that in the cerebrum and spinal cord.
From the whole plant of Frasera tetrapetala PALL. (Gentianaceae), 1, 3-dihydroxy-2, 4, 7-trimethoxyxanthone (I) was isolated as a new compound together with 1-hydroxy-2, 3, 4, 5-tetramethoxyxanthone, oleanolic acid, apigenin, swertisin and methyl 2-hydroxy-3-(1-β-D-glucopyranosyl) oxybenzoate. This result support the Toyokuni's proposal in which this plant could be classified into Frasera rather than Swertia with taxomic evidence.